Introductory Aspects in

Pharmacognosy

Dr. Yousef Abusamra, PhD

Assistant professor of

Pharmacognosy and Phytochemistry

Email: yabusamra@Philadelphia.edu.jo

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ALKALOIDS

• Definition: organic compound which is:

I. With limited distribution in nature.

II. Present in; plant, fungi, bacteria, marine creatures.

III. It has physiological action or poisonous effect.

IV. Biosynthetic pathway amino acid.

Hagnauer system of classification:

A.True alkaloids: contain heterocyclic nitrogen which is

derived from amino acid and is always basic in nature due

to presence of lone pair of electrons on nitrogen.

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B. Proto alkaloids: are simple amines in which nitrogen

is not heterocyclic, and still derived from an amino

acid.

• Example: ephedrine, colchicine, mescaline.

C. Pseudo alkaloids: they are not derived from amino

acids, but they show positive test for alkaloids. (they

may contain heterocyclic nitrogen).

• Examples: purines (caffeine), steroidal and terpenoidal

alkaloids conessine, solanine, protoveratrine, aconine.

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Just to know

(not demanded from the students):

Nearlly, 167 alkaloids were tested, whereas, one-third (35.9%) is

pharmaceutically significant.

There are 5750 different structural skeletons in the total of 21,120

known alkaloids.

It is estimated that 135,500 natural products are derived from

plants, accordingly, alkaloids constitute a percentage of 15.6%.

Represent a vast amount of conformational rigidity to flexibility

(tremendous diversity in chemical structures from linear chains, to

planar, multi-ring systems).

Usually, they are obtained with a high degree of optical purity.

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Why plants produce alkaloids

; Role in the plant

• Venting toxic substances into alkaloids that are

less toxic (detoxification of the poisonous

substances produced during metabolism).

• As a reservoir of nutrients, e.g. storing nitrogen.

• To secure protection against grazing animals due

to the bitter taste and inherent toxicity (most

scientists accept this point more than the other

two).

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Morphine

Aristolochic acid

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MescalineEphedrine

CaffeineConessine

Proto alkaloids:

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Pseudo alkaloids:

Pseudo Alkaloids

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Nomenclature: •According to:

Genus; e.g. Atropine {Atropa belladonna}

Species; e.g. Cocaine; {Erythroxylum coca}

Physiological activity; e.g. Morphine (named after the

Roman god of dreams, Morpheus, who also became the god

of slumber; nap).

Ex: Ergometrine: works on endometrium (uterus lining)

; causes contractions of the uterus to treat heavy vaginal

bleeding after childbirth, (from the ergot)

Discoverer e.g. pelletierine (P. J. Pelletier, French

chemist).

It should generally end with the suffix: ~ine.

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Chemical properties of alkaloids:

• Present in plants in different forms as: salt, ester, N-oxide,

quaternary compound.

N-oxide alkaloids:More water soluble.

Less toxicity.

Less addictive.

• Generally very toxic compounds.

• Have bitter taste.

• A lot of them are unstable compounds in h e a t , l i g h t , p H

changes.

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Physicochemical properties:

Solid crystalline compounds (exception examples:

coniine and nicotine are liquid (they don't have oxygen in

their structure).

Colorless compounds (exception examples: berberine

(yellow), betaine (red).

Sharp melting point because they are pure

compounds in crystal form.

- Can be either 1º, 2º, 3º or 40 alkaloid: Basis of separation;

separatory funnel.

- Basicity depends on the availability of lone pair of

electrons:

1. Electron-donating or electron-withdrawing neighbors.

2. Type of hybridization.

3. Aromaticity.

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Detection of alkaloids:-

1. Wagner's test: (I2/kI): reddish brown precipitate.

2. Mayer’s test: (HgCl2 Creamy precipitate with true

alkaloid.

3. Hagger's test: (Picric acid) Yellow precipitate with

true alkaloid.

4. Dragendroff: (Potassium Bismuth Iodide) reddish

brown precipitate.

5. Tannic acid solution: different alkaloid colored

precipitate .

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Extraction:

-The extraction is fractional extraction (from less polar to

more polar).

Defatting by non-polar solvent (e.g. petroleum ether,

benzene, alkane,….) To get red of chlorophyll, wax,

volatile oil, fixed oil.

-Filtration, for marc, use methanol or ethanol 95%.

-Evaporate by rotary evaporator (to concentrate)

-Add tartaric acid 2% and ethyl acetate. Separate into two

layers:

- Organic layer (for week or neutral alkaloid)

- Aqueous layer (acidic layer, tartaric acid) which has the

alkaloidal salts.

- To break the salt, add NH3 or sodium bicarbonate, then

add ethyl acetate again so will it separate into two layer

again:

- Aqueous layer (quaternary alkaloids 4º)

- Organic layer (for basic alkaloid 10,2º,3º)

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Classification of Alkaloids

N

N

NNH

NH

N

NH

MeN

Tropane

NH

N

N N

NH

Piperidine Pyridine Pyrrolidine Quinoline

Isoquinoline PurineImidazoleIndole

NH

Tetrahydro-

isoquinoline

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Skeletal structures of alkaloids found in medicinal plants.Numbers; I or II refer to the affiliation to the previously mentioned divisions.

Amino Alkaloids• The nitrogen atom of an amino alkaloid is

located in an amino group and is not a

member of a heterocycle, a common feature in

other alkaloids.

Ephedrine:

• Is obtained from Ephedra sinica and related

species.

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• Ephedrine is a sympathomimetic, which means that its effects are similar to those which arise on the stimulation of sympathomimetic nerves.

• Ephedrine causes increased blood pressure and pulse, contraction of blood vessels, and dilation of the bronchi.

• It stimulates the central nervous system like amphetamine, but less strongly.

• Ephedrine sulphate and chloride are used as bronchodilators in the treatment of asthma and colds.

Biosynthesis: is derived from the amino acid phenylalanine.

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Ephedra:

• The dried aerial parts of Ephedra sinica (Gnetaceae) العلندة ، العليق,ذنب الخيل

• Comes from china where it has been used for treatment of common cold.

• Main sources nowadays are India and Pakistan.

• Collected in autumn.

• Pseudoephedrine is a sterioisomer of ephedrine.

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• Pseudoephedrine has α-adrenergic activity and used in treatment of rhinitis because of its ability to decrease swelling of the mucous membranes.

Khat:

• Is the leaf of Catha edulis (Celastraceae) الحرابيةالفصيلة

• A shrub or a small tree native to East Africa. It is cultivated in the high lands of Yemen, Ethiopia and Kenya.

• Leaves are chewed or used for preparation tea, that is drunk as a stimulant.

• Fresh leaves are preferred.

• Khat counteracts fatigue, facilitates strenuous muscular work and causes a light elation (pleasure) with talkativeness and sociability.

• Effect after chewing appears after about half an hour.

• The effects described above are due to cathinone. Its effect is similar to that of amphetamine (Structural similarity)

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Mescaline:

• Is a hallucinating alkaloid which is obtained from the cactus Lophophora williamsii (Cactaceae) الصباريات

• Oral doses (100-400mg) cause euphoria, changes in the conception of time, and brilliant and colored hallucinations.

• Is used to induce models of psychoses in studying the etiology of mental diseases.

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Amphetamine Cathinone

• Biosynthesized from the amino acid, tyrosine.

Capsaicin:

• Is a pungent substance occurring in the fruits of certain

Capsicum species (Solanaceae) which causes irritation

of the skin (rubefacient= redness-causing; increase

circulation).

• Biosynthetically, it originates from phenylalanine.

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Colchicine {Tropolone Alkaloids}:

• Is an alkaloid that can be obtained from the autumn crocus

(the meadow suffron), Colchicum automnale لحلاح , or Gloriosasuperba = flame Lilly (both from the family Colchicaceae).

• The molecule consists of an aromatic group (A) with three

methoxyl groups , a 7-membered ring (B) carrying an

acetylated amino group, and a tropolone ring (C) whose

hydroxyl group is methylated.

• As the nitrogen atom is part of an amide functional group,

colchicine is non-basic.

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• In medicine, colchicine is used as a remedy against gout, a disease

caused by the disposition of u r i c a c i d in the joints.

• It is highly poisonous, and the treatment must be carefully supervised.

• It inhibits division of animal cell, but it is too poisonous to be used to

arrest tumor growth.

B i o s y n t h e s i s :

Ring A is derived from phenylalanine, which also contributes

carbon atoms 5, 6 and 7 of ring B.

The tropolone is derived from tyrosine by ring expansion involving

-carbon atom of tyrosine which becomes C-12 of the tropolone ring.

The nitrogen atom in colchicine is also derived from tyrosine.

The methoxyl substituents come from methionine.

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The tuber has been used in the folkloric medicine for suppression of gout.

The alkaloid is used orally for treatment of gout.

Colchicine induces P O L Y P L O I D Y {{ Polyploid cells and organisms are those containing more than two paired (homologous) sets of chromosomes. Most species whose cells have nuclei (Eukaryotes) are diploid, meaning they have two sets of chromosomes—one set inherited from each parent}}

(so used: to multiply the number of chromosomes in other plants to obtain larger flowers, pollen grains and stomata and also to increase the amount of alkaloids).

Used for familial Mediterranean fever.

Crude drug:

Colchicum seed from the autumn crocus which has an unusual flowering period that gives it its name; in August and September.

It is a European plant, and seeds are produced in Poland, Yugoslavia and Netherlands.

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Polyploidy

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Aziridine alkaloids

• Aziridines are the nitrogenous analogues of epoxides.

• The aziridine group is a 3-membered heterocyclic with one amine group and two methylene groups.

• The simplest compound is aziridine, which is present in many natural products with anticancer and antibacterial.

Mitomycin C:

• Mitomycin C is produced by Streptomyces lavendulae {a species of bacteria from the genus Streptomyces. It is isolated from soils globally and is known for its production of medically useful biologically active metabolites}.

• It is a highly toxic antibiotic with antineoplastic properties which is used for treatment of solid tumors of the bladder, breast, cervix, eye, stomach, and prostate.

• It is a prodrug that requires activation by enzymes such as [1] DT-diaphorase which is an enzyme that targets novel anti-cancer drugs and reduces them for sake of activation, or by the enzyme [2] NADH cytochrome c reductase.

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Aziridine alkaloids

• The mechanism of action involves interaction of the aziridine

ring with guanine in DNA causing intra- and inter-strand DNA

cross-linking, leading to selective inhibition of DNA synthesis,

mutagenesis, induction of DNA repair and induction of

apoptosis {programmed cell death}.

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GuanineMitomycin c

See the next slide

DNA structure

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Aziridine alkaloids

• The compound is unique as an anti-cancer drug in that the

activation preferentially occurs in the hypoxic regions of solid

tumors.

• The main adverse effects are:

1. Delayed cumulative bone marrow suppression, and doses

have to be adjusted according to the effect on the bone

marrow.

2. Renal damage.

3. Pulmonary toxicity.

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Tropane Alkaloids

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Solanaceous Tropane Alkaloids

Occurrence:They are mainly found in Solanaceae family, but also in

Erythroxylaceae ----- كوكيات

Main Alkaloids are:

1- Atropine.

2- Hyoscyamine.

3. Hyoscine (Scopolamine).

Found in the leaves of:

a. Atropa belladonna.

b. Datura stramonium.

c. Hyoscymus spp. like:

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Hyoscyamus niger (البنج الأسود, henbane, black henbane or stinking nightshade) and Hyoscyamus muticus ( .which is indigenous to Egypt (Solanaceae) (البنج المصري

• These plants are toxic and their berries of belladonnaare lethal.

• They are esters of tropic acid and alcoholic basetropanol which have either α- or β- configuration.

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H. muticusH. niger Datura stramonium

M. officinarum (Solanaceae) is limited to small areas of

northern Italy and the coast of former Yugoslavia.

M. autumnalis, the autumn mandrake is native to the

Medditerranean countries, like Palestine, Jordan, Tunisia,

Turkey, Lebanon, Syria, Morocco, … etc.

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d.MandrakeMandragora officinarum & Mandragora

autumnalisتفاحة الشيطان - بيض المجانين - تفاح المجانين

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Mandrake and

witchcraft

Hyoscine (Scopolamine)- Hyoscine is an ester of l -tropic acid with

scopoline base (A tropanol with an oxygen).

- Hyoscine is a syrupy liquid.

(-)-Scopolamine =(-)-Hyoscine

Hyoscyamine

Tropic acidScopoline

EsterEster

Tropic acid

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Atropine =

(-)-hyoscyamine

• Atropine does not exist in nature but is obtained upon

the extraction of hyoscyamine to levo and dextro

racemers which, both, give atropine.

• Atropine, thus, is a mixture of (+) and (-)-hyscyamaine.

α-tropanol (tropane-3 α-ol) gives atropine, while β-tropanol

gives pseudoatropine.

• D and l: polarizes the light domain to the right and left,

respectively. So, atropine is optically is inactive.

Tropane can be considered a combination of piperidine

and pyrrolidine (Pyrrolidine is derived from pyrrolinium cation which is

formed from ornithine in the same way as in the biosynthetic pathway of

nicotine).

α-tropanol

β-tropanol

Hyoscyamine:

** is the major natural alkaloid with negative optical

rotation (l- form) [(-)-hyoscyamine that is racemized

to atropine].

** Because the α-carbon is asymmetric, so two

sterioisomers are possible (racemers).

** During extraction hyoscyamine racemizes to the

optically inactive dl Atropine.

** Both alkaloids are composed of tropine base and

tropic acid.

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β-tropinolα-tropinol

SAM: S-adenosyl methionine

phenylalanine

From

pyrrolinium

cation

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Alkaloids in the form of HCl salts

1- Alkalinize by NaHCO3 pH 7.5

2- Extract with Ether

Ether

Hyoscine free base

(pKa = 6.2)

Aqueous layer

Atropine & Hyoscyamine HCl

(pKa = 9.3)

Convert to oxalate salts,

Fractional Crystallization

(Acetone/ Ether)

Atropine Oxalate

CrystalsHyoscyamine Oxalate

Solution

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Principle: difference in

basicity

Principle: difference in solubility in a

mixture of acetone and

ether